NZ240826A - Hydroxy-substituted pyrrolidine derivatives for controlling diseases caused by parasitic nematodes - Google Patents

Abstract

PCT No. PCT/GB91/02111 Sec. 371 Date Aug. 30, 1983 Sec. 102(e) Date Aug. 30, 1993 PCT Filed Nov. 28, 1991 PCT Pub. No. WO92/09202 PCT Pub. Date Jun. 11, 1992.The use of the compound 2R,5R-dihydroxymethyl-3R,4R-dihydroxypyrrolidine (DMDP) <IMAGE> or an acid addition salt thereof in controlling diseases caused by parasitic nematodes in plants or mammals.

Description

New Zealand Paient Spedficaiion for Paient Number £40826 2408 2 6 : . ' ■ 2, i "2.. • rip | tw,^ .2.; li.-.'vi,! | r'-«- iv1- . 2 5 NOV 1993 po. j. . J31o-. « « ,<*v ■— i~ V 'Pi r. ■' ;. " S W ^'u ■'*'u ^ ^ u - ^ ^ NEW ZEALAND PATENTS ACT, 1953 No.: Date: M.Z. PATfTfvT 0~P' -2 DEC 1991 RECEIVED COMPLETE SPECIFICATION substitution of app,.:c,v<: 4^""ooNn -yccWr>olog^ 41 Gr°^P OL OF PARASITIC NEMATODES (A) We, NATIONAE-^eSE^GFfc©BVEL0PM'ENT ^RP0R?>31€aara British corporation established by Statute of 101 Newington Causeway, London SE1 6BU, England, hereby declare the invention for which we pray that a patent may be granted to us, and the method by which it is to be performed, to be particularly described in and by the following statement:- -l- (followed by page 1A) 2408 2 6 - 1A- 133992 CONTROL OF PARASITIC NEMATODES (A) FIELD OF INVENTION This Invention relates to the control of diseases caused by parasitic nematodes In plants and mammals.

PRIOR ART 05 Since the early 1940s many chemical compounds active against plant parasitic nematodes have been available. These have often displayed undesirable toxic effects, for example the fumlgant dibromochloropropane was withdrawn from the market in 1977, as it was thought to cause sterility in workers. During the 1960's 10 fumigant type nematicides were largely superseded by granular systemic nematicides. These have been in use since then, a representative compound being oxamyl. These compounds are mainly oxlmecarbamates or organophosphate derivatives, and because of their toxicity have to be used in a strictly controlled manner. 15 Accordingly it would be of benefit to have anti-nematode agents that are environmentally favourable, I.e. being non-toxic themselves and in their degradation products to non-target organlsms.

Additional prior art 1s referred to in a separate section after 20 "Summary of the invention", without which Its context would not be clear.

SUMMARY OF THE INVENTION The present Invention provides the use of the compound 2R,5R-di hydroxymethy1-3R,4R-d1hydroxypyrrol1d1ne (DMDP) or an acid addition salt thereof, for use 1n controlling diseases caused by parasitic nematodes in plants. Including crops, and 1n mammals. The Invention also Includes seeds, dressed, coated or impregnated with DMDP or a said salt thereof. 7 •] 0 8 2 6 The mechanism through which DMDP controls diseases caused by parasitic nematodes in plants may include any nematotoxic, nematostatic or anti-feedant effect on either adult or juvenile nematodes, inhibition of hatching of larval forms of nematodes, 05 inhibition of root gall formation by nematode feeding, and further extends to any effect on a nematode that prevents Its acquisition and/or transmission of plant viruses.

DMDP is of natural origin and has been shown to display low phytotoxlcity.

ADDITIONAL PRIOR ART The discovery and extraction of DMDP is described by L. E. Fellows and G. W. 0. Fleet in "Alkaloid Glycosidase Inhibitors from Plants" (In "Natural Products Isolation", G. H. Wagman and R. Cooper, Eds., Elsevier, Amsterdam, 1988, 15 pp 540-565). In that review certain properties of DMDP, Including 1nsect1cidal and insect deterrent activity, both as determined experimentally in feeding tests, are referred to. They are more clearly described in L. E. Fellows, Chemistry in Britain pp 842-844 (1987). These and other properties of DMDP are more extensively 20 reviewed in Chapter 11 of "Plant Nitrogen Metabolism", Plenum Publishing Corporation, 1989, pp 395-427, by L. E. Fellows £i il., especially at pages 410 (which refers to S. V. Evans £±a1., Entomol. Exp. Appl. il, 257-261 (1985), 411 (which refers to the authors' own work and to W. M. Blaney £i al., Entomol. Exp. Appl. 25 26, 209-216 (1984) and 415. See also L. E. Fellows si al-. In "Swalnsonine and Related Glycosidase Inhibitors", L. James, A. D. El be1n, R. J. Molyneux and C. D. Warren, Eds., Iowa State University Press, 1989, pp 396-416. The properties of DMDP referred to therein are not Indicative of an anti-nematode effect. 30 DESCRIPTION OF THE PREFERRED EMBODIMENTS A further advantage of DMDP lies in its mode of application when treating plants, especially crops. Many existing anti-nematode compounds are applied to the soil by broadcasting and Incorporated using rotary cultivation. DMDP can be applied to the leaves, 35 which, somehow produces an anti-nematode action 1n the roots of the 2408 plant. Possibly DMDP is translocated through the phloem, but this is not certain. Hence, DMDP may be applied in the form of a foliar spray Instead of or in addition to the above-mentioned conventional means of application. A suitable dosage for soil application of 05 DMDP is from at least 24 to at most 48 kg/ha at 20 cm depth. DMDP may also be applied by pre-treating plant seeds before sowing.

DMDP is water-soluble and can therefore be applied without a surfactant or dispersing agent. The preferred concentration of active ingredient and rate of application depend on the mode of 10 application and type of effect desired, e.g. they may differ for nematotoxicity and for inhibition of virus transmission. For foliar spraying it is suggested that normally the plants be sprayed with a solution containing 0.01 to 3.5 g./litre, preferably 0.01 to 1.0 g./litre of the active Ingredient, until the spray runs off. 15 Lower concentrations can be more useful in some circumstances, while higher concentrations will often be tolerable.

DMDP displays its properties against a wide range of nematodes affecting plants, e.g. root-knot nematodes, cyst nematodes and virus-transmitting nematodes. Of particular note 1s its activity 20 against the crop-damaging nematodes of the following genera: Meloldogyne, Globodera, Heterodera, Radopholus. Pratvlenchus. H1rschmann1e1la. Scutellonema. Hel icotvlenchus. Tvlenchus. Rotvlenchus. Ditvlenchus. Lonaidorus. Xlphinema. With regard to nematodes which infest mammals, DMDP is active against a wide range 25 of helminthic nematodes, especially those of the following genera: Haemonchus. Teladorsaglft, Nematodirus, Trlchostronavlus. D1 ctvocaulus and Cooperla. particularly the species Haemonchus contortus and Teladorsaala circumclncta (previously classified as Ostertaala circumcincta).

DMDP may be extracted from Derris el 1iptica Benth (Leguminosae) as described by A. Welter £l al (Phytochem., 1976, JL5, 747-749) or may be synthesized from D-glucose (Fuhrman £± &L., Nature, 1984, 307. 755-758); G. W. J. Fleet and R. W. Smith Tetrahedron Letters Z& (11) 1465-1468 (1985) or from L-sorbose (P. Card fil al., J. Org. 35 Chem., 1985, 891-893). 2 4 0 3 2 The above description of DMDP applies also to Its acid addition salts, which can be any which are compatible with the Intended use, e.g. agriculturally or veterinarily acceptable If the use 1s on plants or non-human animals respectively. Such salts can be made 05 In the conventional way from the free base.

The following Examples illustrate the invention. "Tween" 1s a Registered Trade Mark. The units "ppm" signify a solution containing mg. of test compound per litre of water, in solutions for in vitro tests or in solutions for application to leaf 10 surfaces. In the Examples, "DMDP" means the free base.

Example 1 Virus acquisition and transmission experiments The effect of a chemical on virus acquisition by a nematode vector was tested by exposing virus-free nematodes to a virus 15 Infected source plant In the presence of the test chemical. By comparing subsequent rates of virus transmission between treated and untreated nematodes the efficacy of the chemical can be determi ned.

Whether a chemical affects the transmission of the virus can be 20 determined by applying the chemical after the nematodes have acquired the virus, at the time they are about to feed on receptor plants.

Experiments were performed In 25 cm^ plastic pots maintained 1n temperature controlled cabinets (Taylor & Brown, Nematol. medit.. 25 1974, 2, 171-175) using three week old seedlings of Petunia hvbrlda Vllm. The nematode/virus combination used was Xlohlnema dlverslcaudatum vectoring Arabis Mosaic Virus.

Petunia seedlings were potted In 22 ml of 3:1 sand/loam mixture. Forty-eight hours later the plants were inoculated with 30 virus. After a further 24 hours 5 adult nematodes were added to each pot. (The test chemicals are added at this time If virus acquisition is being tested.) There were 10-15 replicates of each treatment. After 4 weeks the nematodes were extracted, and then added to the soil 1n which virus-free receptor plants were 35 growing. (If virus transmission 1s being tested, the test 2408 2 6 chemicals are added at this time.) After a further 4 weeks the nematodes were again extracted and counted. The galls on the roots of the receptor plants were counted, the roots macerated and the sap applied to the leaves of Chenopodium auinoa plants (virus 05 indicators).

Twelve days later the £. qulnoa plants were examined for the symptoms of the virus. There were 10-15 replicates of each treatment in both virus tests. In all cases controls were run 1n which no chemicals were added.

The chemicals tested were DMDP (15 and 30 ppm) and a conventional nematotoxic compound oxamyl (7 ppm).

Table la shows the effect of DMDP inhibiting root gall formation and per cent virus acquisition as compared to the control value. Table lb shows the effect of DMDP inhibiting root gall formation 15 and per cent virus transmission as compared to the control value.

TABLE la Feeding and acquisition of Arabis Mosaic Virus by XlDhinema dlverslcaudatum Treatment Mean No. % virus No. of galls/root acquisition Replicates Control 1.5 33 DMDP 15 ppm 0.5 (66%) 27 (18%) DMDP 30 ppm 0.4 (74%) 7 (79%) 14 Oxamyl 7 ppm 0.3 (80%) 0 (100%) ( ) 1s % reduction in treatment compared to control 2408 2 6 TABLE lb Feeding and transmission of Arabis Mosaic Virus by Xiphinema diversicaudatum Treatment Mean No. % virus No. of gal 1s/root transmi ssion replIcates Control 1.5 64 11 DMDP 15 ppm 0.4 (74%) 72 ( 0%)* DMDP 30 ppm 0.5 (66%) CM 00 11 Oxamyl 7 ppm 0.7 (53%) 1 (98%) 11 ( > 1s % reduction in treatment compared to control * treatment values higher than control Example 2 Hatch Test The hatch test examines the effect of the test chemicals on the egg hatch of Globodera pallida, the white Potato Cyst Nematode 05 (PCN).

Ten PCN cysts of uniform size and colour were put 1n a tube with 0.25 ml of the test compound solution (concs. 50 ppm and 100 ppm) and 0.75 ml of potato root diffusate. Root dlffusate normally stimulates the juveniles to hatch from eggs 1n the cysts. There 10 were 4 replicates of each treatment. Twice each week the liquid was removed and the number of hatched live and dead juveniles counted. The diffusate/chemlcal mixture was replenished after each nematode count. The tubes were stored at 19°C between counts.

Table 2a shows the number of hatched juveniles, dead or alive, 15 as the means from four replicates. The same data are also expressed as % effect. This Table shows that DMDP greatly decreases the number of juveniles hatching from cysts.

This experiment was repeated using Globodera rostochlensis. Table 2b shows the % decrease 1n nematodes alive as compared to the 20 control after 4 weeks. From Table 2b, it can be seen that DMDP provides better effects than Its acid salt. 2408 2 TABLE 2a Potato Cvst Nematode Hatch Test Treatments Hatched Juveniles Total Juveniles Live Dead Hatched (% increase)* (1 increase)* (1 decrease)* days exposure Control 698 16 714 DMDP 50 ppm 374 (46) 68 (325) 442 (38) DMDP 100 ppm 203 (71) 91 (468) 294 (59) 24 days exposure Control 1257 32 1289 DMDP 50 ppm 1056 (16) 112 (250) 1168 ( 9) DMDP 100 ppm 601 (52) 150 (368) 751 (42) TABLE 2b Globodera rostochiensis cvst Hatch Test Test Compound Cone (ppm) 220 100 50 12.5 6.25 3.12 DMDP 32 38 52 52 41 0 DMDP. HC1 0 0 0 0 27 31 21 * All percentages are based on the control value Example 3 In Vitro Toxicity Test Groups of ten active adult Xiphinema diversicaudatum were hand-picked Into Individual watchglasses containing distilled water. At a given time the batches of nematodes were transferred Into 1 ml aliquots of test compound, at various concentrations of the test compound, or for the control Into 1 ml of distilled water. There were three replicates of each treatment. At two 2 4 0 8 2 6 intervals, viz. 48 and to 72 hours, the number of nematodes which were immobilised were recorded. They were considered as immobile If they failed to move when stimulated by prodding with a bristle. All tests were carried out at 5°C. 05 Table 3a shows the in vitro toxicity of DMDP over a range of concentrations. The percent immobility shown 1s corrected for control immobilities using Abbott's formula. Note the decrease in in vitro toxicity at 200 ppm and above. There is also an anomalous drop in toxicity at 25 ppm.

In similar tests differences in toxicity to adult and juvenile nematodes were found. Table 3b shows the EC50 values (effective concentration required to Immobilise 507. of the total number of nematodes) calculated from the results.

This experiment was repeated, replacing dlverslcaudatum 15 with Globodera rostochlensls. These results are shown in Table 3c, from which it can be seen that both DMDP and its acid salt are toxic to nematodes.

Table 3a In vitro toxicity (adult Xiphinema diversicaudatum) Test compound Cone (ppm) 10 50 100 200 Percent Immobility 500 DMDP 48 hrs 15 72 hrs 39 11 35 0 9 63 78 4 0 0 Table 3b In vitro toxicity EC50 values (ddm) (XiDhinema dlverslcaudatum) Test compound Nematode stage tested Test duration 48 hrs 72 hrs DMDP Adult DMDP Juvenile 87.0 44.0 94.0 0.08 2408 2 6 Table 3c In vitro toxicity (Globodera rostochiensis) Test Compound Cone (ppm) 2.5 50 100 DMDP 37 44 50 37 DMDP.HC1 88 56 50 50 50 Example 4 Table 4 shows the dose-dependent activity of DMDP, using three tests: the split-pot experiment, the mini-pot experiment and the gall test experiment. 05 a. Split-pot test The test shows whether the anti-nematode agents of the invention have a repellent or antifeedant effect on the nematodes and/or a nematicidal effect.

A 'split-pot', i.e. a pot divided Into two sections by a 10 fine mesh material (see Alphey fii al. Revue Nematol. 1988, 11(4), 399-404), was used. Each side was filled with 37 ml of soil (3:1 sand:loam mixture). Test compounds at the concentrations shown in Table 4 were added to the soil on the side 1n which a Petunia seedling had been planted. To the other side 100 adult Xiphinema 15 dlverslcaudatum were added. There were 8 replicates of each treatment.

After 21 days the two halves of the pot were separated and the nematodes were extracted from the soil 1n each half. Root galls were recorded on plants from the treated sides (Table 20 4a(D). The numbers of live and dead nematodes from each half were counted and are shown 1n Table 4a(ii).

Table 4a(i) shows that DMDP has an antifeedant action against nematodes at all concentrations tested. Table 4a(ii) shows that 80 ppm DMDP also possesses a nematotoxic effect in 25 that on the plant side more nematodes were immobilised than 1n the pot to which oxamyl was applied. 240 b. Mini-pot test This test identifies the nematicidal effect of the chemical in soil and its effect on nematode feeding behaviour.

Petunia seedlings were planted in 22 ml of soil 05 (sandrloam - 3:1). The test compound solution or water (control) with 5 or 10 adult Xiphinema diversicaudatum were added to the soil. There were 10 replicates for each treatment. After 3 weeks the nematodes were extracted and the number of galls induced by nematode feeding on the roots were recorded and 10 expressed as a mean per cent reduction of the control value.

Table 4b shows that DMDP has a nematode repellent or antifeedant action. The most effective rate of DMDP was 25 ppm. c. Gall test In the gall test, tomato seedlings, stimulated to produce 15 fine adventitious roots by removing the main root system, were planted in tubes containing 25 g of fine, sieved dry sand, 350 Meloidoovne incognita (J2) and DMDP, in solution in water. The effect of DMDP on the ability of the nematodes to gall the plant roots was studied over a 10-12 day period. A water control 20 was included in the test. There were 10 replicates of each treatment.

Table 4c shows the results, from which 1t will be seen that DMDP 1s equally effective in the range 2.5-25 ppm but less effective at 50 and 240 ppm. The various tests Indicate similar 25 levels of activity of DMDP used between 2.5 ppm and 100 ppm Table 4 4a.(1) Split-pot Experiment (X- d1versicaudatum/Petunia) Chemical/conc (ppm) Mean reduction galls/root as % of control DMDP/15 DMDP/30 DMDP/80 63 83 89 2408 26 - li - 4a-C11> Mean numbers of nematodes recovered after 21 days 1n the planted and non-planted sides of the split pot (X- dlversicaudatum/Petunla) Total Mobile Immobile Nematodes Nematodes Nematodes Test conc Chemical <ppm) Plant No plant Plant No plant Plant No plant DMDP 16 27 24 3 DMDP 32 24 14 21 11 3 3 DMDP 80 12 11 13 4 Oxamyl 17 21 13 14 4 7 Control _ 33 16 31 12 2 4 4b. Mini-pot Experiment (X- diversicaudatum/Petunia) Chemical/conc (ppm) Mean reduction galls/root as % of control nematodes/pot 10 nematodes/pot DMDP/8 70 DMDP/14 70 DMDP/25 94 72 DMDP/50 72 83 DMDP/100 65 100 4c. Gall Test (£L incognita/Tomato) Chemical/conc (ppm) Reduction in galls/root as % of control DMDP/2.5 76 DMDP/12.5 70 DMDP/25 72 DMDP/50 50 DMDP/240 47 2408 2 6 Example 5 Mode of Application a) root application To test whether the anti-nematode agent would be more 05 effective when taken up systemically by plants, the mini-pot test was adapted. The roots of Petunia hvbrida were removed and the cut ends of the stems from which the newly formed roots were growing were put in a solution of test compound (concentration as shown in Table 5) for 24 hours prior to the start of the experiment. The 10 effects of these treated plants to X. diversicaudatum were compared to that of plants whose cut ends had been immersed in water for 24 hours. Table 5 shows that root uptake following soil application Is a suitable method of treatment with DMDP. b) foliar application The mini-pot test and gall test described 1n Example 4 were repeated but the test compounds were administered by being painted on to the leaves of the tomato seedlings. In these tests, 0.4 ml test compound in solution In water at 200 ppm, or water alone, together with 0.05% "Tween 80" wetting solution, were painted onto 20 the leaves.

The reductions in galling of 86% in the m1n1-pot test and 79% In the gall test, over the controls, show that the effect of the test compounds was expressed in the root system to provide protection against nematodes.

TABLE 5 Activity following uptake through root - details as in text Mini-pot test: Petunla/XIphinema diverslcaiidatum <21 days) Chemical/conc (ppm) % reduction In root galling relative to controls Oxamyl/50 DMDP/15 DMDP/30 DMDP/100 92 83 100 58 2408 2 6 Example 6 Phytotoxicity data DMDP was tested on three different plant species at 200 ppm for 14 days using methods outlined in the min1-pot test. The 05 seedlings were then left to grow for 16 days and the % growth measured relative to control plants. Root length and shoot length were also measured.

Table 6 shows the effect of DMDP on plant growth. All figures are % growth relative to controls (100% = same as control, 10 >100% = greater than control).

Rye grass when treated with DMDP only grew to 65% of the control weight. This may not be significant in the field as the concentration of DMDP (200 ppm) used was twice its effective dosage required to control nematodes.

TABLE 6 Phytotoxicity data (all at 200 ppm soil water) Root length Shoot length Total weight Chemical TOM OSR RG TOM OSR RG TOM OSR RG Oxamyl 107 84 108 91 95 93 103 104 107 DMDP 90 98 105 90 97 74 100 100 65 Plants TOM = Tomato (cv. Moneymaker) 0SR = Oilseed rape (cv. Bienvenue) RG - Rye grass (cv. Melie) Example 7 Canister test Small 60ml clear canisters were filled with approximately 25g soil. 1ml test compound and 1ml water containing 1500 PCN eggs was 20 added. Small pieces of Deslree potato with sprout were placed Into the compost. Lids pierced 3-4 times were used to close the canisters. The canisters were then put on a tray, covered with black polythene and kept at a constant 20*C. After 4 weeks the first cyst count was taken, then every following week until the end °4Q8 26 of the eighth. Table 7 shows the % reduction in cysts, as compared to the control. It can be seen that DMDP was effective in reducing the number of cysts developing.

Table 7 Canister test (Globodera rostochiensis) 05 t reduction i n cysts Test Compound Conc (ppm) 3.12 6. .26 12.5 25 50 100 200 DMDP 7 0 14 46 43 35 7 DMDP.HC1 0 0 0 7 7 43 0 Example 8 Methods of Application II As an extension to Example 5, further experimentation was undertaken in sand and soil, or a variety of plants and nematodes 10 to demonstrate the different methods of applying DMDP. 8(1) Sand Drench Test in a Tube Glass tubes (7.5cm x 2.5cm) were filled with 24.5g sieved dried sand. 4ml nanopure water was added and a hole made 1n the sand, lml test compound and 1ml water containing 350 Meloldoavne iavanica 15 were added Immediately before a tomato seedling was planted 1n the hole. All tubes were then left for 14 days. In this experiment and in 8(2) below, seedlings were prepared by having their roots cut off and fine adventitious roots allowed to regenerate prior to use. Table 8(1) shows the effect of DMDP and its acid salt over a 20 range of concentrations. Results are shown as % reduction in live nematodes as compared to a control (no test compound). 8(2) Sand Foliar Test in a Tube 3 glass tubes (7.5cm x 2.5cm) were filled with 24.5g sieved dried sand. 5ml nanopure water was added and a tomato seedling 25 planted 1n the tube. Non-absorbant cotton wool was Inserted around the base of the seedling to protect the sand from the test chemical to be sprayed. The tubes were placed 1n an incubator overnight. 2 4 0 8 2 Next day, each plant was sprayed with O.lml test chemical from an airbrush and returned to the incubator. On the following day, 1ml water containing 350 Meloidoovne javanica was added to each tube. All tubes were then left for 14 days. Table 8(2) shows the effect 05 of DMDP and its acid salt on a range of plants. Results are shown in % as in Table 8(1). 8(3) Foliar Application 2.5cm pots were filled with 75g of Levington universal and sand in a 3:1 ratio. Tomato plants (34 days old) were planted 1n these 10 pots and 1ml of water added. The soil was protected with filter paper and the pots left overnight in a glasshouse. Next day, each plant was sprayed with 0.3ml test compound from an airbrush and then left in the glasshouse overnight. Next day the filter paper was removed and 350 Meloidoavne javanica or Meloidoovne Incognita 15 1n 1ml water were added to the soil. The pots were then left for 12 days after which the number of live and dead nematodes were counted. Table 8(3) shows the effect of DMDP on a) Meloidoovne javanica and b) Meloidoovne Incognita. 8(4) Soil Apd!iration The procedure of 8(3) was repeated, except that on the first day, 1ml test compound and 1ml water with nematodes were added to the soil and the pots left for 14 days. Results are shown 1n the usual manner in Table 8(4).

Table 8(1) Sand Drench % reduction In galling by id. lavanlca Test Compound Conc (ppm) 200 100 50 25 10 5 1 DMDP.HC1 DMDP (Expt. 1) DMDP (Expt. 2) 47 51 30 18 43 13 77 72 79 76 56 57 53 56 68 63 71

Claims (10)

2408 2 6 - 16 - Table 8(2) Sand Foliar 1 reduction 1n galling by M. iavanica Plant Test Compound Conc (ppm) 3200 2400 1600 800 400 Tomato DMDP 59 0 9 DMDP.HC1 18 5 9 Peppers DMDP 7 7 30 0 DMDP.HC1 9 0 7 0 Aubergines DMDP 38 43 34 9 DMDP.HC1 44 50 19 19 Table 8(3) Soil Foliar % reduction in galling by a) M. iavanica b) M- incognita Test Compound Conc 1600 1000 800 400 200 100 50 25 10 1 0.1 a) DMDP 27 27 22 22 a) DMDP 35 28 22 39 34 b) DMDP 24 24 26 30 31 b) DMDP 23 22 Table 8(4) Soil Drench % reduction in galling by a) ij. iavanica b) M- incognita Test Compound Conc (ppm) 100 50 20 10 1.0 0.1 0.01 DMDP 28 19 21 DMDP 28 30 29 20 8 WHAT I/WE CLAIM IS:

1. The use of the compound 2R,5R-dihydroxymethyl-3R,4R-dihydroxypyrrolidine (DMDP), or an acid addition salt thereof in controlling diseases caused by parasitic nematodes in plants or non-human mammals. 05

2. The use according to Claim 1, wherein the compound is applied to plants or crops in a foliar spray.

3. The use according to Claim 1, wherein the compound is applied to plants through the soil.

4. The use according to Claim 1, 2 or 3 wherein the parasitic 10 nematode attacks plants or crops and is of the genus Meloidoovne. Globodera. or Xiphinema.

5. The use according to Claim 1, wherein the parasitic nematode Infests mammals and is of the genus Haemonchus. Teladorsagia. Nematodirus. Trichostronavlus. Dictvocaulus or Cooperia. 15

6. Seeds dressed, coated or impregnated with a compound specified 1n claim 1.

7. A method of protecting plants from diseases caused by nematodes comprising spraying a compound specified in Claim 1, on the leaves of the said plant. 20

8. A method of protecting plants from diseases caused by nematodes comprising applying a compound specified in Claim 1, to the soi1.

9. The use of the compound as defined in claim 1 substantially as herein described with reference to any example thereof.

10. A method of protecting plants from diseases caused by nematodes as defined in claim 7 or claim 8 substantially as herein described with reference to' any example thereof. .&£.CT!SH . rePrt^y-CGr'i G-gooP o mi-rep 8QCi 1993 By tho authorised agents. A J PARK S SON Per,^Vu.O — Lj